**2. Life cycle of** *L. botrana* **on grapevines**

*L. botrana* is a multivoltine species with a facultative diapause (physiological state of inactivity). The number of generations depends on latitude, photoperiod, humidity, temperature, climate, microclimate and food type [11]. In Europe, two generations per year are common in Germany, Switzerland, Austria and northern France, while three generations (and sometimes four) have been reported in southern France, Spain, Portugal, Greece and Italy [12, 13]. In Chile at least three and possibly four annual generations are known [14].

The eggs of the first generation are deposited separately or in groups of two or three on grapevine buds, pedicels and flowers [15]. Their shape is elliptical, flat and slightly convex, and they measure between 0.65–0.90 mm long by 0.45–0.75 mm wide. Recently laid eggs are translucent and creamy white in color (**Figure 1A**), turning pale yellow with time (**Figure 1B**). They then turn black, with the head of the developing larva visible (**Figure 1C**) [16]. Finally, the egg hatches 7–11 days after laying, depending on temperature and humidity conditions (**Figure 1D**) [8, 15]. Once the larva emerges from the egg, only the shell or the round and nacreous mark of the shell remains (**Figure 1E**).

*L. botrana* larvae have five development stages (**Figure 2**): I (L1: 0.9–1-0 mm), II (L2: 1.9–3.0 mm), stage III (L3: 4.5–5.0 mm), stage IV (L4: 6.0–7.0 mm) and stage V (L5: 10.0–11.0 mm). Larval development concludes after 20 to 30 days in optimal conditions of 26.7°-29.4°C and 40–70% relative humidity [14].

First generation larvae are called the anthophagous generation, since they attack the plant in or near its flowering season, feeding on flower buttons, flowers and occasional small recently formed fruits. First generation larvae form "nests" or glomerules before and during flowering (**Figure 3**) [14]. These glomerules are formed by various flower buds joined together by silk threads spun by the larvae [8]. Damage caused by first generation larvae on the vines have minimal repercussions [17]. However, larvae in the second generation cause decreased vine productivity, since they attack developing grapes, perforating the skin and feeding on their pulp. Finally, these grapes are scared (**Figure 4**), dry out, fall or rot, depending on their size and the ambient humidity. Third generation larvae, by comparison with second generation larvae (both called carpophagous generations) produce greater damage to vine productivity, since the grapes are matured or in the maturation process [14]. Therefore, larval action exposes their sugary juices, favoring the entry,

#### **Figure 1.**

*L. botrana eggs. A, creamy white egg. B, yellow eggs. C, black head egg. D, larva hatching, E) round and nacreous mark.*

*Integrated Pest Management of* Lobesia botrana *with Microorganism in Vineyards… DOI: http://dx.doi.org/10.5772/intechopen.99153*

#### **Figure 2.**

*L. botrana larvae. A, newborn larva. B, young larva. D, mature larva. E, stage V larva spinning a grayishwhite silk cocoon for the pupation process.*

#### **Figure 3.**

L. botrana *glomerules on grape bunches.*

#### **Figure 4.** *Grape damage from* L. botrana *larvae.*

establishment and proliferation of microorganisms responsible for diseases including *Botrytis cinerea* (Persoon: Fries) (*Sclerotiniaceae)* [18] and black *Aspergillus* (*Aspergilus niger* and *Aspergilus carbonarius*) which produces ochratoxin A [1].

*L. botrana* pupae are elongated, with a green to dark brown color. The average length of a male and female pupa is 5.5 mm and 7.0 mm, respectively, while the average width is 1.6–1.7 mm (**Figure 5A**). Males have 4 abdominal segments, and

females have 3. Their eyes, antennae, wings and abdominal segments can be seen in their structure. Pupae are covered by a silky, white, fused and continuous cocoon.

In vineyards, *L. botrana* hibernates in the pupal stage principally beneath the grapevine bark, in trunk cracks, soil and fallen leaves. During this period the pupae are in diapause, presenting a thick, highly hydrophobic cocoon. This tissue protects the pupa from low temperatures and water (**Figure 5B**) [19]. Full pupal development in diapause takes around 90 days while the pupal state during spring and summer is around 12–14 days, or 130°C days [14, 16].

In springtime, when temperatures rise, adults emerge from pupae in diapause. They emerge in stages, beginning before grapevine budding or extending over several weeks. The first adults to emerge are generally males, but in the later part of the flight period females predominate.

Adult *L. botrana* specimens are 6.0–8.0 mm long with a wingspan of 11.0– 13.0 mm. Both sexes have a dorsal design with a cross-sectional band on the front wing pair, which can be seen with the wings laid to rest over the body. Male lack a side fold in their front wings; their back wings are whitish with a brown edge, while female rear wings are completely brown [16]. They can live from one to three weeks. Their activity is crepuscular, remaining inactive during the day and hiding in leaves and bunches. They mate in flight (1 to 6 days after emerging), females generally mate once in their lives. Egg laying begins one or two days after mating, and each female can lay between 80 and 160 eggs [16].

Regarding the dispersion capacity of *L. botrana* moths, males can fly several meters above vegetation and use air currents for longer migrations, while females generally spread over small areas and cannot go beyond 100 m [20]. This indicates that *L. botrana* colonization in new territories occurs mainly due to transferring pest-infested materials.
